Rat FⅨ(Coagulation Factor Ⅸ) ELISA Kit

This ELISA kit applies to the in vitro quantitative determination of Rat F concentrations in serum, plasma and other biological fluids.

Test principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat F. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat F and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat F, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat F. You can calculate the concentration of Rat F in the samples by comparing the OD of the samples to the standard curve.

Assay type

Sandwich

Format

96T

Assay time

4.5h

Reactivity

Rat

Detection Method

Colormetric

Detection Range

0.16 10 ng/mL

Sensitivity

0.10 ng/mL

Sample Volume Required Per Well

100 ¼L

Sample Type

Serum, plasma and other biological fluids

Specificity

This kit recognizes Rat F in samples. No significant cross-reactivity or interference between Rat F and analogues was observed.

Typical data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration(ng/mL)

O.D

Average

Corrected

10

2.298 2.354

2.326

2.249

5

1.539 1.587

1.563

1.486

2.5

0.913 0.875

0.894

0.817

1.25

0.414 0.45

0.432

0.355

0.63

0.301 0.271

0.286

0.209

0.32

0.193 0.165

0.179

0.102

0.16

0.119 0.139

0.129

0.052

0

0.069 0.085

0.077

--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat F¨ were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat F¨ were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision

Inter-assay Precision

Sample

1

2

3

1

2

3

n

20

20

20

20

20

20

Mean (ng/mL)

0.50

1.54

4.87

0.48

1.53

4.50

Standard deviation

0.03

0.09

0.23

0.03

0.07

0.24

C V (%)

6.00

5.84

4.72

6.25

4.58

5.33

Recovery

The recovery of Rat F¨ spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type

Range (%)

Average Recovery (%)

Serum (n=5)

89-102

96

EDTA plasma (n=5)

91-106

98

Cell culture media (n=5)

88-100

95

Linearity

Samples were spiked with high concentrations of Rat F¨ and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5)

EDTA plasma (n=5)

Cell culture media (n=5)

1:2

Range (%)

93-108

98-114

96-109

Average (%)

98

106

101

1:4

Range (%)

88-101

85-100

83-94

Average (%)

93

92

87

1:8

Range (%)

88-99

85-100

84-96

Average (%)

93

92

91

1:16

Range (%)

91-105

83-95

84-95

Average (%)

98

90

90

Kit components & Storage

An unopened kit can be stored at 4„ƒ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.